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Biosafety
Organisms We Used
We use E. coli K-12 MG1655 and phage M102 in our project. They help us to control the number of S. mutans, pathogen in the mouth. As we hope to use these organisms inside our mouth, we do biosafety research, and design suicide circuits in E. coli to prevent it from causing disease when dropping out of our mouth. We will discuss biosafety, laboratory safety respectively.
Strain information of E. coli K-12 MG1655
We use E. coli K-12 MG1655 and phage M102 in our project. They help us to control the number of S. mutans, pathogen in the mouth. As we hope to use these organisms inside our mouth, we do biosafety research, and design suicide circuits in E. coli to prevent it from causing disease when dropping out of our mouth. We will discuss biosafety, laboratory safety respectively.
Strain information of bacteriophage M102
E. coli K-12 is in risk group 1, which is the most harmless. It is not able to colonize human’s intestinal tract under normal conditions, thus it won’t cause disease in human[1].
E. coli suicide circuit design
Strain information of bacteriophage M102 Phage M102 belongs to the family of Siphoviridae with morphotype B1[2], which is tail long and noncontractile, head isometric. Bacteriophage M102 is specific to S. mutans, there is little concern of risks to human and environment.
Lab safety
We conducted our experiments in Biosafety level 1 laboratory, which is suitable for work involving well-characterized agents not known to consistently cause disease in healthy adult humans, and of minimal potential hazard to laboratory personnel and the environment according to CDC(Centers for Disease Control and Prevention in United states). Furthermore, professors checked our experiments before we did it. And we have written the protocols to confirm that we had right procedures.
Lab training
Every person in NYMU iGEM team has 3 in 1 training directed by professors, who led us step by step. Thus, some toxic chemicals are emphasized, like DNA dye used in electrophoresis. After doing all steps overseen by professors, all people could use instruments properly.
Protection
Everyone must wear coat and gloves before entering the laboratory with long pants and shoes which covered our feet entirely. If we need to go out of the experimental zone, we take off the coat and gloves, and washed hands with antibacterial hand wash, not to touch door knob with gloves. We have done this strictly, so to prevent taking the bacteria out of lab.
Waste
There is chemical and bacteria mixture after conducting the experiment, we gather then into a bottle. Clean it daily to prevent explosion because of air made by bacteria. After autoclaving the waste, we pour it with a lot of water to dilute the toxic chemicals. Instead of carcinogen EtBr, we used safer dyes to reduce contamination.
Reference
- Escherichia coli K-12 Derivatives Final Risk Assessment , TSCA (http://epa.gov/biotech_rule/pubs/fra/fra004.htm)
- van der Ploeg, J. R. (2007). Genome sequence of Streptococcus mutans bacteriophage M102. FEMS Microbiol Lett 275, 130–138.
